Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
J Virol Methods. 2012 May;181(2):164-9. doi: 10.1016/j.jviromet.2012.02.003. Epub 2012 Feb 21.
Avian influenza viruses (AIVs) and Newcastle disease viruses (NDVs) co-circulate in the poultry population in China. These viruses cause repeated disease outbreaks that exhibit similar clinical symptoms and epidemiological patterns. H5 and H9 influenza viruses are the major pathogens infecting poultry stocks. Recently, H3 AIV (one of the main subtypes in waterfowl) has become endemic in chickens. A multiplex reverse-transcriptase polymerase chain reaction (mRT-PCR) assay was designed for simultaneous detection and differentiation of avian H3, H5, H9 subtype AIVs and NDVs. Four primer sets were evaluated, three of which specifically targeted the hemagglutinin genes of H3, H5 and H9 AIVs, while the other targeted the NDV fusion gene. The sensitivity and specificity of the mRT-PCR assay was determined. The assay detected the major clades or genotypes of all of the reference AIVs and NDVs currently circulating in China. In addition, the mRT-PCR results obtained from screening 380 clinical swabs and 12 experimental tracheal samples were consistent with those obtained using conventional virus isolation methods. The mRT-PCR assay was established successfully for the detection and differentiation of avian H3, H5, and H9 subtype AIVs and NDVs. The method should, therefore, provide a valuable diagnostic tool for these infections.
禽流感病毒（AIVs）和新城疫病毒（NDVs）在中国的家禽群体中共同循环。这些病毒导致了反复发作的疾病爆发，表现出相似的临床症状和流行病学模式。H5 和 H9 流感病毒是感染家禽的主要病原体。最近，H3 AIV（水禽中的主要亚型之一）已在鸡中成为地方性流行。设计了一种多重逆转录聚合酶链反应（mRT-PCR）检测方法，用于同时检测和区分禽 H3、H5、H9 亚型 AIVs 和 NDVs。评估了四个引物组，其中三个特异性针对 H3、H5 和 H9 AIV 的血凝素基因，而另一个针对 NDV 的融合基因。确定了 mRT-PCR 检测方法的敏感性和特异性。该检测方法可检测到目前在中国流行的主要 AIVs 和 NDVs 的分支或基因型。此外，通过对 380 份临床拭子和 12 份实验性气管样本进行筛查，mRT-PCR 结果与常规病毒分离方法一致。成功建立了用于检测和区分禽 H3、H5 和 H9 亚型 AIVs 和 NDVs 的 mRT-PCR 检测方法。因此，该方法应为这些感染提供一种有价值的诊断工具。
